在对大鼠肝脏核糖体进行紫外线照射后，通过沉降分离出与28-S和18-S rRNA交联的核糖体蛋白。

Ribosomal proteins cross-linked to 28-S and 18-S rRNA separated by sedimentation after ultraviolet irradiation of rat-liver ribosomes.

作者信息

Uchiumi T, Terao K, Ogata K

出版信息

Eur J Biochem. 1983 May 16;132(3):495-9. doi: 10.1111/j.1432-1033.1983.tb07388.x.

DOI:10.1111/j.1432-1033.1983.tb07388.x

PMID:6852010

Abstract

Rat liver 80-S ribosomes, irradiated with 254 nm ultraviolet light for 15-60 min (0.63-2.5 X 10(5) quanta/ribosome), were treated with sodium dodecyl sulfate; 18-S and 28-S RNA fractions were isolated by sucrose density-gradient centrifugation. The protein moieties of the 18-S and 28-S rRNA fractions were labeled with 125I. After digestion of the rRNA fractions with RNase A and T1, the cross-linked proteins were analyzed by two-dimensional acrylamide gel electrophoresis and then autoradiography. Cross-linked proteins were examined further by dodecyl sulfate/acrylamide gel electrophoresis of individual radioactive spots of proteins and their proteolytic peptides on two-dimensional gels. The results showed that S2 and S15 were cross-linked to 18-S rRNA, and L5, L6 and L8 to 28-S rRNA. Similar results were obtained when isolated 40-S or 60-S subunits were irradiated.

摘要

用波长254nm的紫外线照射大鼠肝脏80-S核糖体15 - 60分钟（0.63 - 2.5×10⁵量子/核糖体），然后用十二烷基硫酸钠处理；通过蔗糖密度梯度离心分离出18-S和28-S RNA组分。用¹²⁵I标记18-S和28-S rRNA组分的蛋白质部分。在用核糖核酸酶A和T1消化rRNA组分后，通过二维聚丙烯酰胺凝胶电泳分析交联蛋白，然后进行放射自显影。通过对二维凝胶上蛋白质及其蛋白水解肽的各个放射性斑点进行十二烷基硫酸钠/聚丙烯酰胺凝胶电泳，进一步检查交联蛋白。结果表明，S2和S15与18-S rRNA交联，L5、L6和L8与28-S rRNA交联。当分离的40-S或60-S亚基受到照射时，也获得了类似的结果。